[Histonet] Cresyl echt violet stain

[Rena Fail]

Thank you Sarah

-----Original Message-----
From: Sarah Jones [mailto:SJones <@t> cvm.tamu.edu] 
Sent: Thursday, February 26, 2004 12:18 AM
To: histonet <@t> lists.utsouthwestern.edu; rfail <@t> TOOLKITMAIL.COM
Subject: Re: [Histonet] Cresyl echt violet stain


>>> rfail <rfail <@t> toolkitmail.com> 2/25/2004 8:36:40 AM >>>
I would appreciate it if someone could help me out. I need the cresyl
echt violet procedure from Frieda Carson's book.Thank you


There are two procedures from Carson's book, I've included them both
here.  I've also added a procedure I prefer from Bancroft's book.  
Sarah



Cresyl Echt Violet Method for Nissl Substance
Fixative: 10% neutral buffered formalin, cut paraffin sections at 6-8
microns

Reagents
Stock Cresyl Echt Violet Solution
0.5 gm Cresyl echt violet (C.I. 51010)
80 ml distilled water
20 ml absolute alcohol
warm the distilled water, add the Cresyl echt violet, mix, and then add
the alcohol.

Working Cresyl Echt Violet Solution
45 ml Cresyl echt violet stock solution
15 drops glacial acetic acid

Procedure
1.	Deparaffinize and hydrate the sections to distilled water.
2.	Stain sections in working Cresyl echt violet for 8 minutes.
3.	Dehydrate sections with 95% and absolute alcohol, two changes
each
4.	Clear in two changes of xylene and mount in synthetic resin.

Results:  Nissl substance and nuclei-blue-purple, background-colorless


                        Cresyl echt Violet from LFB procedure 0.1%
Cresyl Echt Violet 0.1 gm Cresyl echt violet 100 ml distilled water Just
before use, add 15 drops of 10% acetic acid solution, filter, and
preheat.  
This solution is not very stable.

Procedure
1.	Deparaffinize sections and hydrate to distilled water.
2.	Place in Cresyl echt violet solution for 6 minutes. Filter and
preheat Cresyl echt violet solution to 57                 degrees C.
just before use.  Keep hot during staining.
3.	Differentiate in several changes of 95% alcohol.
4.	Dehydrate in absolute alcohol, clear and mount.

Results:  Nissl substance and nuclei-violet

Ref. Histotechnology, A Self-Instructional Text, by Freida Carson, pages
160, 161, 175


and now for something completely different...................... ;) I
personally prefer this procedure from Theory and Practice of
Histological Techniques, by John Bancroft and Alan Stevens, page 353 &
354

Cresyl Violet Acetate for Nissl Substance

Fixation: Formalin fixed tissue              Sectioning: Paraffin
sections cut from 6-15 microns

Solutions:

0.1% Cresyl Violet Stain
0.1 gm Cresyl Violet Acetate (Sigma Catalog No. C-5042) 
100 ml Distilled water
Dissolve the dye in water.  Add 0.25 ml of Glacial Acetic Acid to each
100 ml of dye. 
This stain is stable for years. (until you get your tombstone shaped
sticker from ASCP)

Cresyl Violet Differentiator
44 ml 95% ethanol
60 ml chloroform
9 drops of Glacial Acetic Acid

Technic:
1.  Deparaffinize sections and hydrate to distilled water.
2.  Stain in 0.1 % Cresyl Violet Stain for 5 minutes.
3.  Wash in tap water.
4.  Differentiate in Cresyl Violet Differentiator, dipping until color
no longer runs off.  This will take 5-10 dips. 5.  Check differentiation
under the microscope. (Nissl and nuclei only should be stained) 6.
Dehydrate, clear and mount.

Results:  Nissl substance-blue




Sarah Jones HT(ASCP)
Dept. of Vet. Anatomy & Public Health
Histology Lab
Texas A&M University
College Station, TX 77843-4458
phone: 979-845-3177
fax:  979-458-3499